The nucleocapsid protein of SARS coronavirus has a high binding affinity to the human cellular heterogeneous nuclear ribonucleoprotein A1.
The nucleocapsid protein of SARS coronavirus has a high binding affinity to the human cellular heterogeneous nuclear ribonucleoprotein A1.
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SARS冠状病毒的Nucleocapsid蛋白与人类细胞异质性核核糖核蛋白A1具有高的结合亲和力。
DOI：
10.1016/j.febslet.2005.03.080
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发表时间：
2005-05-09
期刊：
影响因子：
3.5
通讯作者：
Jiang H
中科院分区：
文献类型：
--
作者：
Luo H;Chen Q;Chen J;Chen K;Shen X;Jiang H
SARS冠状病毒（SARS_CoV）的核衣壳蛋白(N)是病毒粒子的主要结构成分，是参与病毒RNA复制和翻译的多功能蛋白。异质核核糖核蛋白A1 （hnRNP A1）与细胞核中的前mRNA剪接和细胞质中的翻译调控有关。本报告通过相关的生物物理和生化分析，发现SARS_CoV的核衣壳蛋白（SARS_N）与人类hnRNP A1具有高结合亲和力。GST下拉结果清楚地表明，SARS_N蛋白可以在体外直接特异性结合人hnRNP A1。酵母双杂交实验进一步表明，这种结合与SARS_N的片段（aa 161-210）和hnRNP A1的Gly - rich结构域（aa 203-320）有关。此外，表面等离子体共振（SPR）技术的动力学分析表明，SARS_N蛋白与人hnRNP A1具有特异性的结合亲和力，K D值为0.35±0.02 μM （K on = 5.83±0.42 × 103 M−1 s−1,K off = 2.06±0.12 × 10−3 s−1）。提示SARS_N和hnRNP A1蛋白可能都在SARS_CoV复制/转录复合体内，SARS_N/human hnRNP A1相互作用可能在SARS_CoV RNA合成调控中起作用。此外，测定结果表明，SARS_N蛋白只有一个与人hnRNP A1相互作用的结合域，这与小鼠肝炎病毒（MHV）的结合情况不同，MHV的核衣壳蛋白（MHV_N）有两个参与MHV_N/hnRNP A1相互作用的结合域，这表明SARS_N蛋白与MHV_N相比，可能通过不同的结合方式与人hnRNP A1结合，以进一步发挥其功能。
The nucleocapsid (N) protein of SARS coronavirus (SARS_CoV) is a major structural component of virions, which appears to be a multifunctional protein involved in viral RNA replication and translation. Heterogeneous nuclear ribonucleoprotein A1 (hnRNP A1) is related to the pre‐mRNA splicing in the nucleus and translation regulation in the cytoplasm. In this report, based on the relevant biophysical and biochemical assays, the nucleocapsid protein of SARS_CoV (SARS_N) was discovered to exhibit high binding affinity to human hnRNP A1. GST pull‐down results clearly demonstrated that SARS_N protein could directly and specifically bind to human hnRNP A1 in vitro. Yeast two‐hybrid assays further indicated in vivo that such binding relates to the fragment (aa 161–210) of SARS_N and the Gly‐rich domain (aa 203–320) of hnRNP A1. Moreover, kinetic analyses by surface plasmon resonance (SPR) technology revealed that SARS_N protein has a specific binding affinity against human hnRNP A1 with K D at 0.35 ± 0.02 μM (k on = 5.83 ± 0.42 × 103 M−1 s−1 and k off = 2.06 ± 0.12 × 10−3 s−1). It is suggested that both SARS_N and hnRNP A1 proteins are possibly within the SARS_CoV replication/transcription complex and SARS_N/human hnRNP A1 interaction might function in the regulation of SARS_CoV RNA synthesis. In addition, the determined results showed that SARS_N protein has only one binding domain for interacting with human hnRNP A1, which is different from the mouse hepatitis virus (MHV) binding case where the nucleocapsid protein of MHV (MHV_N) was found to have two binding domains involved in the MHV_N/hnRNP A1 interaction, thereby suggesting that SARS_N protein might carry out a different binding mode to bind to human hnRNP A1 for its further function performance in comparison with MHV_N.
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DOI：
10.1016/s0140-6736(03)13077-2
发表时间：
2003-04-19
期刊：
Lancet (London, England)
影响因子：
--
作者：
Peiris JS;Lai ST;Poon LL;Guan Y;Yam LY;Lim W;Nicholls J;Yee WK;Yan WW;Cheung MT;Cheng VC;Chan KH;Tsang DN;Yung RW;Ng TK;Yuen KY;SARS study group
通讯作者：
SARS study group
影响因子：
3.8
作者：
Nelson, GW;Stohlman, SA;Tahara, SM
通讯作者：
Tahara, SM
影响因子：
2.9
作者：
Huang, QL;Yu, LP;Olejniczak, ET
通讯作者：
Olejniczak, ET
影响因子：
5.6
作者：
Cartegni, L;Maconi, M;Biamonti, G
通讯作者：
Biamonti, G
影响因子：
5.7
作者：
Xu, RM;Jokhan, L;Krainer, AR
通讯作者：
Krainer, AR